Metabolic pathway rerouting in Paraburkholderia rhizoxinica evolved long-overlooked derivatives of coenzyme F420
2019
Coenzyme F 420 is a specialized redox cofactor with a highly negative redox potential. It supports biochemical processes like methanogenesis, degradation of xenobiotics or the biosynthesis of antibiotics. Although well-studied in methanogenic archaea and actinobacteria, not much is known about F 420 in Gram-negative bacteria. Genome sequencing revealed F 420 biosynthetic genes in the Gram-negative, endofungal bacterium Paraburkholderia rhizoxinica , a symbiont of phytopathogenic fungi. Fluorescence microscopy, high-resolution LC-MS, and structure elucidation by NMR demonstrated that the encoded pathway is active and yields unexpected derivatives of F 420 (3PG-F 420 ). Further analyses of a biogas-producing microbial community showed that these derivatives are more widespread in nature. Genetic and biochemical studies of their biosynthesis established that a specificity switch in the guanylyl transferase CofC re-programmed the pathway to start from 3-phospho-D-glycerate, suggesting a rerouting event during the evolution of F 420 biosynthesis. Furthermore, the cofactor activity of 3PG-F 420 was validated, thus opening up perspectives for its use in biocatalysis. The 3PG-F 420 biosynthetic gene cluster is fully functional in E. coli , enabling convenient production of the cofactor by fermentation.
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